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1.
Arch Immunol Ther Exp (Warsz) ; 70(1): 27, 2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36318344

RESUMO

In particular conditions, inhibition of an immune response is required to prevent tissue damage. Among these conditions are diseases caused by an over-reactive immune response, such as autoimmune or allergic disorders, or imminent organ rejection after transplantation. To avoid tissue damage, drug-mediated systemic immune suppression is an option, but it comes with high costs in the form of susceptibility to viral and bacterial infections. Thus, the induction of antigen-specific tolerance is preferable. Extracellular vesicles (EVs) are capable of delivering antigen together with immunosuppressive signals and may be used to specifically induce antigen-specific tolerance. However, naturally occurring EVs are heterogeneous and not all of them show immunosuppressive character. In our trials to engineer cell culture derived EVs to increase their tolerogenic potential, we equipped them with immunosuppressive miRNA mimics. Small EVs (sEVs) were isolated and purified from the human monocytic THP-1 cell line or from healthy donor peripheral blood mononuclear cells, and electroporated with miR-494 and miR-146a mimics. The acquired immunosuppressive potential of the modified sEVs was demonstrated by their ability to alter the major histocompatibility complex molecules and co-stimulatory receptors present on dendritic cells (DCs). To avoid allogeneic responses, the same cells that produced the sEVs served also as recipient cells. In contrast to the treatment with unmodified sEVs, the tolerogenic sEVs impeded lipopolysaccharide-induced maturation and kept DCs in a more immature developmental stage. Our experiments show that simple manipulations of sEVs using immunosuppressive cargo can lead to the inhibition of DC maturation.


Assuntos
Vesículas Extracelulares , MicroRNAs , Humanos , MicroRNAs/metabolismo , Células Dendríticas , Leucócitos Mononucleares , Vesículas Extracelulares/metabolismo , Diferenciação Celular , Imunossupressores/farmacologia , Antígenos/metabolismo
2.
Int J Mol Sci ; 22(11)2021 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-34199901

RESUMO

The establishment of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) technology for eukaryotic gene editing opened up new avenues not only for the analysis of gene function but also for therapeutic interventions. While the original methodology allowed for targeted gene disruption, recent technological advancements yielded a rich assortment of tools to modify genes and gene expression in various ways. Currently, clinical applications of this technology fell short of expectations mainly due to problems with the efficient and safe delivery of CRISPR/Cas9 components to living organisms. The targeted in vivo delivery of therapeutic nucleic acids and proteins remain technically challenging and further limitations emerge, for instance, by unwanted off-target effects, immune reactions, toxicity, or rapid degradation of the transfer vehicles. One approach that might overcome many of these limitations employs extracellular vesicles as intercellular delivery devices. In this review, we first introduce the CRISPR/Cas9 system and its latest advancements, outline major applications, and summarize the current state of the art technology using exosomes or microvesicles for transporting CRISPR/Cas9 constituents into eukaryotic cells.


Assuntos
Sistemas CRISPR-Cas , Vesículas Extracelulares/genética , Edição de Genes , Técnicas de Transferência de Genes , Terapia Genética/métodos , Animais , Humanos
3.
Anal Chem ; 93(13): 5476-5483, 2021 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-33769802

RESUMO

Extracellular vesicles (EVs) have attracted considerable interest due to their role in cell-cell communication, disease diagnosis, and drug delivery. Despite their potential in the medical field, there is no consensus on the best method for separating micro- and nanovesicles from cell culture supernatant and complex biological fluids. Obtaining a good recovery yield and preserving physical characteristics is critical for the diagnostic and therapeutic use of EVs. The separation of a single class of EVs, such as exosomes, is complex because blood and cell culture media contain many nanoparticles in the same size range. Methods that exploit immunoaffinity capture provide high-purity samples and overcome the issues of currently used separation methods. However, the release of captured nanovesicles usually requires harsh conditions that hinder their use in certain types of downstream analysis. A novel capture and release approach for small extracellular vesicles (sEVs) is presented based on DNA-directed immobilization of antiCD63 antibody. The flexible DNA linker increases the capture efficiency and allows for releasing EVs by exploiting the endonuclease activity of DNAse I. This separation protocol works under mild conditions, enabling the release of vesicles suitable for analysis by imaging techniques. In this study, sEVs recovered from plasma were characterized by established techniques for EV analysis, including nanoparticle tracking and transmission electron microscopy.


Assuntos
Exossomos , Vesículas Extracelulares , Nanopartículas , Sistemas de Liberação de Medicamentos , Fenômenos Magnéticos
4.
Int J Mol Sci ; 21(12)2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32549407

RESUMO

The ability of exosomes to transport different molecular cargoes and their ability to influence various physiological factors is already well known. An exciting area of research explores the functions of exosomes in healthy and pathological pregnancies. Placenta-derived exosomes were identified in the maternal circulation during pregnancy and their contribution in the crosstalk between mother and fetus are now starting to become defined. In this review, we will try to summarize actual knowledge about this topic and to answer the question of how important exosomes are for a healthy pregnancy.


Assuntos
Exossomos/fisiologia , Feto/citologia , Placenta/citologia , Feminino , Humanos , Troca Materno-Fetal , Gravidez
5.
Curr Pharm Des ; 25(42): 4464-4485, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31808383

RESUMO

BACKGROUND: Exosomes open exciting new opportunities for advanced drug transport and targeted release. Furthermore, exosomes may be used for vaccination, immunosuppression or wound healing. To fully utilize their potential as drug carriers or immune-modulatory agents, the optimal purity of exosome preparations is of crucial importance. METHODS: Articles describing the isolation and purification of exosomes were retrieved from the PubMed database. RESULTS: Exosomes are often separated from biological fluids containing high concentrations of proteins, lipids and other molecules that keep vesicle purification challenging. A great number of purification protocols have been published, however, their outcome is difficult to compare because the assessment of purity has not been standardized. In this review, we first give an overview of the generation and composition of exosomes, as well as their multifaceted biological functions that stimulated various medical applications. Finally, we describe various methods that have been used to purify small vesicles and to assess the purity of exosome preparations and critically compare the quality of these evaluation protocols. CONCLUSION: Combinations of various techniques have to be applied to reach the required purity and quality control of exosome preparations.


Assuntos
Exossomos/química , Exossomos/fisiologia , Sistemas de Liberação de Medicamentos , Humanos , Ácidos Nucleicos , Proteínas
6.
Cells ; 8(7)2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31269655

RESUMO

BACKGROUND: Cancer-induced immunosuppression is antigen-specific rather than systemic and the mechanisms for the antigen specificity are incompletely understood. Here we explore the option that tumor-associated antigens (TAAs) may be transferred to antigen-presenting cells (APCs), together with immunosuppressive molecules, through cancer-derived small extracellular vesicles (sEVs), such as exosomes. Stimulation of a suppressive phenotype in the very same APCs that take up TAAs may yield antigen-specific tolerance. METHODS: sEVs isolated from patient-derived or well-established melanoma cell lines were used to demonstrate the transfer of major histocompatibility complex (MHC) molecules to the surface of APCs. The immunosuppressive influence of sEVs was assessed by flow cytometry analysis of activation markers, cytokine expression, and mixed lymphocyte reactions. RESULTS: MHC class I molecules were transferred from melanoma cells to the cell surface of APCs by sEVs. Concomitantly, CD86 and CD40 co-stimulatory molecules were down-regulated and IL-6 production was strongly induced. TGF-ß transported by sEVs contributed to the promotion of a suppressive phenotype of APCs. CONCLUSION: The presented results indicate the existence of a hitherto undescribed mechanism that offers an explanation for antigen-specific tolerance induction mediated by cancer-derived sEVs.


Assuntos
Células Apresentadoras de Antígenos/imunologia , Antígenos de Neoplasias/imunologia , Vesículas Extracelulares/imunologia , Melanoma/imunologia , Evasão Tumoral/imunologia , Antígenos de Neoplasias/metabolismo , Linhagem Celular Tumoral , Vesículas Extracelulares/metabolismo , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , Melanoma/patologia
7.
8.
Anticancer Res ; 37(12): 6779-6789, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29187456

RESUMO

BACKGROUND/AIM: MicroRNAs (miRNAs) transported in melanoma-derived exosomes function as intercellular messengers supporting tumor survival and progression. Hypoxia increases melanoma phenotypic plasticity, drug resistance, and metastasis. MATERIALS AND METHODS: We determined the miRNA profiles in exosomes derived from melanoma cells grown under hypoxic and normoxic conditions by microarray analyses and reverse transcription-polymerase chain reaction (RT-PCR) in order to analyze the potential influence of vesicle-transported miRNAs on cancer-related pathways and transcriptional programs. RESULTS: Despite phenotypical differences of the four cell lines used, their exosomes shared the majority of miRNAs. The levels of three miRNAs were higher in normoxic exosomes, whereas 15 miRNAs were significantly more abundant under hypoxic conditions. Pathway analysis pointed at several cellular processes contributing to proliferation, drug resistance, and modification of the tumor microenvironment, including immunosuppression. CONCLUSION: The miRNA-expression profiles of exosomes from patient-derived melanoma cells are modified by oxygen concentration and reflect the phenotypic changes of melanoma cells under different growth conditions.


Assuntos
Exossomos/genética , Perfilação da Expressão Gênica/métodos , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Hipóxia Celular , Linhagem Celular Tumoral , Humanos , Melanoma/genética , Melanoma/patologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
9.
Arch Immunol Ther Exp (Warsz) ; 65(4): 311-323, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28101591

RESUMO

Extracellular vesicles, including exosomes, constitute an important element of intercellular communication by carrying a variety of molecules from producer to target cells. The transport of mRNA and miRNA can directly modulate gene expression in the target cells. The miRNA content in exosomes is characteristic for the cell from which the vesicles were derived enabling the usage of exosomes as biomarkers for the diagnosis various diseases, including cancer. Cancer-derived exosomes support the survival and progression of tumors in many ways and also contribute to the neutralization of the anti-cancer immune response. Exosomes participate in all known mechanisms by which cancer evades the immune system. They influence the differentiation and activation of immune suppressor cells, they modulate antigen presentation, and are able to induce T-cell apoptosis. Although cancer-derived exosomes mainly suppress the immune system and facilitate tumor progression, they are also important sources of tumor antigens with potential clinical application in stimulating immune responses. This review summarizes how exosomes assist cancer to escape immune recognition and to acquire control over the immune system.


Assuntos
Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Exossomos/metabolismo , Vesículas Extracelulares/metabolismo , Imunoterapia/métodos , Neoplasias/metabolismo , Animais , Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Terapia de Imunossupressão , MicroRNAs/genética , Neoplasias/patologia , Evasão Tumoral
10.
J Drug Target ; 20(5): 389-400, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22424091

RESUMO

BACKGROUND: Guanosine (G)-rich DNA and RNA sequences can adopt a defined secondary structure, the G-quadruplex, which consists of multiple stacked G-tetrads. Each G-tetrad has four Gs arranged in a planar configuration and held together by hydrogen bonding. G-quadruplexes are found in chromosomal DNA and RNA transcripts, particularly in telomeric sequences, and in regulatory regions of many genes including oncogenes. PURPOSE: This review summarizes how G-quadruplexes can be employed for anticancer therapies and discusses possible mechanisms. METHODS: The Medline database was searched using the terms "G-rich oligonucleotide (GRO)", "G-tetrad", and "G-quadruplex". RESULTS: Drugs which bind to and stabilize G-quadruplexes can be employed to suppress the elongation of telomers and the gene transcription and translation of oncogenes. G-quadruplex stabilization results in senescence and apoptosis of cancer cells. Besides long-chain nucleic acids, also GRO are able to acquire G-quadruplex conformation to build up a variety of stable structures. Selected aptamers show a highly specific binding capacity to their target molecules, similar to antibodies. Some GRO were shown to induce cell death, preferentially in cancer cells; they demonstrated remarkable anticancer activity in preclinical and first clinical studies. CONCLUSION: G-quadruplexes can be both, targets and tools in anticancer drug development.


Assuntos
Antineoplásicos/farmacologia , Quadruplex G , Neoplasias/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Desenho de Fármacos , Humanos , Neoplasias/patologia , Oncogenes/genética , Telômero/metabolismo , Transcrição Gênica/efeitos dos fármacos
11.
Int J Occup Med Environ Health ; 24(4): 348-58, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22038570

RESUMO

OBJECTIVES: The aim of this study was to investigate the cyto- and genotoxicity of nanocomposites (NCs) and generation of reactive oxygen species (ROS) as a result of particle-cell interactions. MATERIALS AND METHODS: Titanium dioxide (TiO(2)-Ag) and ion-exchange resin (Res-Ag), both coated with silver (Ag), were examined. The murine macrophage J774A.1 cells were incubated in vitro with NC at different concentrations for 24 h. Cytotoxicity was analyzed by the methylthiazolyldiphenyl-tetrazolium bromide reduction test (MTT reduction test). ROS generation was assessed by incubation of cells with dichlorodihydrofluorescein diacetate (DCF) and flow cytometry. DNA damage was detected by comet assay and included single-strand breaks (SSB), alkali-labile sites (ALS) and oxidative DNA damage after formamidopyrimidine glycosylase (FPG) treatment. The tail moment was used as an indicator of DNA damage. RESULTS: TiO(2)-Ag was not cytotoxic up to 200 µg/ml, whereas IC(50) for Res-Ag was found to be 23 µg/ml. Intracellular ROS levels were elevated after 4 h of exposure to Res-Ag at the concentration of 50 µg/ml. Both types of NC induced fragmentation of DNA strands, but only one of the composites caused damage to purine bases. TiO(2)-Ag induced SSB of DNA at concentrations of 10 and 5 µg/ml. For Res-Ag, a concentration-dependent increase in tail moments was observed. CONCLUSIONS: Silver-coated nanocomposites (both TiO(2)-Ag and Res-Ag) may cause genotoxic effects in murine macrophages J774A.1. Res-Ag increased generation of ROS which suggested that toxicity of Res-Ag in murine macrophages is likely to be mediated through oxidative stress. This paper will support industry and regulators alike in the assessment of hazards and risks and methods for their mitigation at the earliest possible stage in material and product development.


Assuntos
Citotoxinas/análise , Citotoxinas/intoxicação , Mutagênicos/análise , Mutagênicos/intoxicação , Nanoestruturas/intoxicação , Prata/efeitos adversos , Prata/metabolismo , Indústria Têxtil , Bioensaio/métodos , Citotoxinas/genética , Dano ao DNA/efeitos dos fármacos , Humanos , Exposição Ocupacional/efeitos adversos , Exposição Ocupacional/análise , Polônia , Espécies Reativas de Oxigênio/análise
12.
Cell Mol Life Sci ; 68(24): 4023-32, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21833586

RESUMO

Oxidatively damaged biomolecules impair cellular functions and contribute to the pathology of a variety of diseases. RNA is also attacked by reactive oxygen species, and oxidized RNA is increasingly recognized as an important contributor to neurodegenerative complications in humans. Recently, evidence has accumulated supporting the notion that tRNA is involved in cellular responses to various stress conditions. This review focuses on the intriguing consequences of oxidative modification of tRNA at the structural and functional level.


Assuntos
Modelos Genéticos , Estresse Oxidativo , RNA de Transferência/química , Metabolismo Energético , Oxirredução , RNA de Transferência/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Enxofre/química
13.
Melanoma Res ; 20(1): 21-34, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19949351

RESUMO

Metastatic melanoma is a highly life-threatening disease. The lack of response to radiotherapy and chemotherapy highlights the critical need for novel treatments. Parthenolide, an active component of feverfew (Tanacetum parthenium), inhibits proliferation and kills various cancer cells mainly by inducing apoptosis. The aim of the study was to examine anticancer effects of parthenolide in melanoma cells in vitro. The cytotoxicity of parthenolide was tested in melanoma cell lines and melanocytes, as well as melanoma cells directly derived from a surgical excision. Adherent cell proliferation was measured by tetrazolium derivative reduction assay. Loss of the plasma membrane integrity, hypodiploid events, reactive oxygen species generation, mitochondrial membrane potential dissipation, and caspase-3 activity were assessed by flow cytometric analysis. Microscopy was used to observe morphological changes and cell detachment. Parthenolide reduced the number of viable adherent cells in melanoma cultures. Half maximal inhibitory concentration values around 4 mumol/l were determined. Cell death accompanied by mitochondrial membrane depolarization and caspase-3 activation was observed as the result of parthenolide application. Interestingly, the melanoma cells from vertical growth phase and melanocytes were less susceptible to parthenolide-induced cell death than metastatic cells when drug concentration was at least 6 mumol/l. Reactive oxygen species level was not significantly increased in melanoma cells. However, preincubation of parthenolide with the thiol nucleophile N-acetyl-cysteine protected melanoma cells from parthenolide-induced cell death suggesting the reaction with intracellular thiols as the mechanism responsible for parthenolide activity. In conclusion, the observed anticancer activity makes parthenolide an attractive drug candidate for further testing in melanoma therapy.


Assuntos
Melanoma/tratamento farmacológico , Sesquiterpenos/farmacologia , Neoplasias Cutâneas/terapia , Tanacetum parthenium/química , Acetilcisteína/farmacologia , Adulto , Animais , Antineoplásicos/antagonistas & inibidores , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Adesão Celular/efeitos dos fármacos , Processos de Crescimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Interações Medicamentosas , Feminino , Citometria de Fluxo , Humanos , Masculino , Melanoma/metabolismo , Melanoma/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Pessoa de Meia-Idade , Espécies Reativas de Oxigênio/metabolismo , Sesquiterpenos/antagonistas & inibidores , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia
14.
Br J Haematol ; 148(6): 868-78, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19995395

RESUMO

One characteristic of chronic lymphocytic leukaemia (CLL) lymphocytes is high expression of CD23, which has previously been identified as a downstream target for NOTCH2 signalling. The mechanisms regulating NOTCH2-dependent CD23 expression, however, are largely unknown. This study showed that peripheral CLL cells overexpressed transcriptionally active NOTCH2 (N2(IC)), irrespective of their prognostic marker profile. When placed in culture, NOTCH2 activity was spontaneously decreased in 25 out of 31 CLL cases (81%) within 24 h. DNA-bound N2(IC) complexes could be maintained by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) or by gamma-interferon (IFN-gamma), two CLL characteristic inducers of CD23 expression. Inhibition of PKC-delta by RNA interference or by rottlerin antagonised PMA-induced NOTCH2 activation and also suppressed NOTCH2 activity in CLL cases with constitutively activated NOTCH2 signalling. In 23 out of 29 CLL cases tested (79%), DNA-bound N2(IC) complexes were found to be resistant to the gamma-secretase inhibitor (GSI) DAPT, suggesting that GSIs will be only effective in a subset of CLL cases. These data suggest that deregulation of NOTCH2 signalling is critically involved in maintaining the malignant phenotype of CLL lymphocytes and point to a link between PKC-delta and NOTCH2 signalling in the leukemic cells.


Assuntos
Leucemia Linfocítica Crônica de Células B/imunologia , Proteínas de Neoplasias/metabolismo , Proteína Quinase C-delta/metabolismo , Receptor Notch2/metabolismo , Receptores de IgE/metabolismo , Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Humanos , Interferon gama/imunologia , Leucemia Linfocítica Crônica de Células B/genética , Prognóstico , Proteína Quinase C-delta/antagonistas & inibidores , Interferência de RNA , Receptor Notch2/genética , Transdução de Sinais , Acetato de Tetradecanoilforbol/imunologia , Células Tumorais Cultivadas
15.
Biochem Pharmacol ; 70(10): 1431-42, 2005 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-16185667

RESUMO

New derivatives of daunorubicin (DRB), doxorubicin (DOX), and epidoxorubicin (EDOX) with an amidine group bonded to C-3' of daunosamine moiety with either morpholine or hexamethyleneimine ring attached to the amidine group are studied in this paper. We have shown that all of these newly synthesized anthracycline derivatives inhibit human leukemia K562 cell line proliferation but only some of them induce erythroid differentiation when used at subtoxic concentrations. Morpholine derivative of DOX has the greatest potential to inhibit proliferation and to induce differentiation in vitro. The correlation between these two cellular processes was also significant for other tested compounds. In cell cycle analysis, we have demonstrated that those anthracycline derivatives that exert the greatest cytostatic potential caused G(2)/M arrest, which in turn, might contribute to the development of a differentiating phenotype. The concentrations of the compounds used in the study are pharmacologically relevant. These new potent inducers of differentiation might be exploited as anticancer drugs for treatment of leukemia by differentiation therapy.


Assuntos
Antraciclinas/toxicidade , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células K562 , Antraciclinas/química , Antraciclinas/farmacocinética , Antineoplásicos/química , Antineoplásicos/toxicidade , Tamanho Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Células Eritroides/citologia , Células Eritroides/efeitos dos fármacos , Células Eritroides/metabolismo , Feminino , Fase G2/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/genética , Humanos , Antígeno Ki-67/efeitos dos fármacos , Antígeno Ki-67/genética , Microscopia de Varredura por Sonda/métodos , Morfolinas/química , Morfolinas/toxicidade
16.
J Gene Med ; 4(3): 282-91, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12112645

RESUMO

BACKGROUND: Somatic gene therapy requires safe and efficient techniques for the gene transfer procedure. The ovine mammary gland is described as a model system for the evaluation of somatic gene transfer methods. METHODS: Different gene delivery formulations were retrogradely injected into the mammary gland of lactating sheep. The efficiency of the gene transfer was subsequently measured by the detection of the secreted transgene products in the milk. To counteract the milk flow in the lactating gland caused by the permanent milk production, a newly developed pretreatment of the mammary gland with hyperosmotic solutions was applied. In addition, in vivo electroporation of DNA into the mammary gland is described. RESULTS: Gene transfer using naked DNA or simple complexes of DNA with polycations did not result in traceable amounts of reporter gene products. However, utilizing the complex cationic lipid DOSPER, a peak expression of about 400 ng/ml was observed 6 days after transfection. Maximum expression rates of more than 1 microg/ml were obtained by combining hyperosmotic pretreatment and receptor-mediated gene transfer. For the in vivo electroporation, the proof of principle for this technique in the mammary gland is reported. CONCLUSIONS: The ovine mammary gland turned out to be a very well suited as a model system for evaluation and optimization of various gene transfer protocols.


Assuntos
Técnicas de Transferência de Genes , Lactação , Glândulas Mamárias Animais/metabolismo , Animais , Eletroporação , Ensaio de Imunoadsorção Enzimática , Feminino , Genes Reporter , Glândulas Mamárias Animais/anatomia & histologia , Concentração Osmolar , Ovinos , Soluções , Transfecção
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